Genetic and genomic evaluation of age at first calving in Italian Holsteins.

Age at first calving (AFC) represents the nonproductive period of ∼2 yr in Holstein cows, and thus, it has a relevant effect on the cost of rearing replacements in the dairy herd. In the present study, we aimed to evaluate genetic and genomic aspects of AFC in the Italian Holstein population. Data of 4,206,218 heifers with first calving between 1996 and 2020 were used. Age at first calving averaged 26.09 ± 3.07 mo and decreased across years. Heritability was estimated using a linear animal model which included the fixed effects of herd-year-season of birth and classes of gestation length, and the random animal additive genetic effect fitted to a pedigree-based relationship matrix. The EBV and genomically EBV (GEBV) were obtained, and they were standardized to mean 100 and standard deviation 5, where animals above the mean are those contributing to reduce AFC. Heritability estimates of AFC ranged from 0.031 to 0.045. The trend of sires' GEBV was favorable and indicated a reduced AFC across years. Approximate genetic correlations between GEBV of AFC and GEBV of other economically important traits were calculated on a subset of genotyped females born after 2015. Moderate favorable associations of AFC with production traits (0.39-0.51), udder depth (0.40), interval from first to last insemination in heifer (-0.43), and longevity (0.34) were assessed. Overall, the greatest lifetime productive performances and most favorable days open in first lactation were observed when heifers calved at 22 to 23 mo. In contrast, progeny of sires with GEBV of AFC above the mean yielded more milk, fat, and protein in first lactation, and had shorter days open than progeny of sires with GEBV of AFC below the mean. Results suggested that breeding strategies to improve AFC should be pursued, also considering genetic correlations between AFC and traits which are already part of the Italian Holstein breeding objective. The inclusion of AFC in an aggregate index is expected to contribute to enhance farm income.

Effectiveness of mid-infrared spectroscopy for the prediction of cow milk metabolites.

Proton nuclear magnetic resonance (1H NMR) spectroscopy is acknowledged as one of the most powerful analytical methods with cross-cutting applications in dairy foods. To date, the use of 1H NMR spectroscopy for the collection of milk metabolic profile is hindered by costly and time-consuming sample preparation and analysis. The present study aimed at evaluating the accuracy of mid-infrared spectroscopy (MIRS) as a rapid method for the prediction of cow milk metabolites determined through 1H NMR spectroscopy. Bulk milk (n = 72) and individual milk samples (n = 482) were analyzed through one-dimensional 1H NMR spectroscopy and MIRS. Nuclear magnetic resonance spectroscopy identified 35 milk metabolites, which were quantified in terms of relative abundance, and MIRS prediction models were developed on the same 35 milk metabolites, using partial least squares regression analysis. The best MIRS prediction models were developed for galactose-1-phosphate, glycerophosphocholine, orotate, choline, galactose, lecithin, glutamate, and lactose, with coefficient of determination in external validation from 0.58 to 0.85, and ratio of performance to deviation in external validation from 1.50 to 2.64. The remaining 27 metabolites were poorly predicted. This study represents a first attempt to predict milk metabolome. Further research is needed to specifically address whether developed prediction models may find practical application in the dairy sector, with particular regard to the screening of dairy cows' metabolic status, the quality control of dairy foods, and the identification of processed milk or incorrectly stored milk.

Invited review: Iodine level in dairy products-A feed-to-fork overview.

The theme of iodine in the dairy sector is of particular interest due to the involvement and the interconnection of several stakeholders along the dairy food chain. Iodine plays a fundamental role in animal nutrition and physiology, and in cattle it is an essential micronutrient during lactation and for fetal development and the calf's growth. Its correct use in food supplementation is crucial to guarantee the animal's recommended daily requirement to avoid excess intake and long-term toxicity. Milk iodine is fundamental for public health, being one of the major sources of iodine in Mediterranean and Western diets. Public authorities and the scientific community have made great efforts to address how and to what extent different drivers may affect milk iodine concentration. The scientific literature concurs that the amount of iodine administered through animal feed and mineral supplements is the most important factor affecting its concentration in milk of most common dairy species. Additionally, farming practices related to milking (e.g., use of iodized teat sanitizers), herd management (e.g., pasture vs. confinement), and other environmental factors (e.g., seasonality) have been identified as sources of variation of milk iodine concentration. Overall, the aim of this review is to provide a multilevel overview on the mechanisms that contribute to the iodine concentration of milk and dairy products.

Nuclear magnetic resonance spectroscopy to investigate the association between milk metabolites and udder quarter health status in dairy cows.

Nuclear magnetic resonance spectroscopy was applied to investigate the association between milk metabolome and udder quarter health status in dairy cows. Mammary gland health status was defined by combining information provided by traditional somatic cell count (SCC) and differential SCC (DSCC), which expresses the percentage of neutrophils and lymphocytes over total SCC. Quarter milk samples were collected in triplicate (d 1 to 3) from 10 Simmental cows, 5 defined as cases and 5 defined as controls according to SCC levels at d 0. A total of 120 samples were collected and analyzed for bacteriology, milk composition, SCC, DSCC, and milk metabolome. Bacteriological analysis revealed the presence of mostly coagulase-negative staphylococci in quarter milk samples of cows defined as cases. Nuclear magnetic resonance spectra of all quarter samples were first analyzed using the unsupervised multivariate approach principal component analysis, which revealed a specific metabolomic fingerprint of each cow. Then, the supervised cross-validated orthogonal projections to latent structures discriminant analysis unquestionably showed that each cow could be very well identified according to its milk metabolomic fingerprint (accuracy = 95.8%). The comparison of 12 different models, built on bucketed 1-dimensional NOESY spectra (noesygppr1d, Bruker BioSpin) using different SCC and DSCC thresholds, corroborated the assumption of improved udder health status classification ability by joining information provided by both SCC and DSCC. Univariate analysis performed on the 34 quantitated metabolites revealed lower levels of riboflavin, galactose, galactose-1-phosphate, dimethylsulfone, carnitine, hippurate, orotate, lecithin, succinate, glucose, and lactose, and greater levels of lactate, phenylalanine, choline, acetate, O-acetylcarnitine, 2-oxoglutarate, and valine, in milk samples with high somatic cells. In the 5 cases, results of the udder quarter with the highest SCC compared with its symmetrical relative were in line with quarter-level findings. Our study suggests that increased SCC is associated with changes in milk metabolite fingerprint and highlights the potential use of different metabolites as novel indicators of udder health status and milk quality.

Characterization of the genetic polymorphism linked to the ß-casein A1/A2 alleles using different molecular and biochemical methods.

The 2 major subvariants of ß-casein (A1 and A2), coded by CSN2 gene, have received great interest in the last decade both from the scientific community and the dairy sector due to their influence on milk quality. The consumption of the A1 variant, compared with the A2 variant, has a potential negative effect on human health after its digestion but, at the same time, its presence improves the milk technological properties. The aim of the present study was to compare the best method in terms of time required, costs, and technical engagement for the identification of ß-casein A1 and A2 variants (homozygous and heterozygous animals) in milk to offer a reliable service for large-scale screening studies. Two allele-specific PCR procedures, namely RFLP-PCR and amplification refractory mutation system (ARMS-PCR), and one biochemical technique (HPLC) were evaluated and validated through sequencing. Manual and automated DNA extraction protocols from milk somatic cells were also compared. Automated DNA extraction provided better yield and purity. Chromatographic analysis was the most informative and the cheapest method but unsuitable for large-scale studies due to lengthy procedures (45 min per sample). Both allele-specific PCR techniques proved to be fast and reliable for differentiating between A1 and A2 variants but more expensive than HPLC analysis. Specifically, RFLP-PCR was the most expensive and labor-demanding among the evaluated techniques, whereas ARMS-PCR was the fastest while also requiring less technical expertise. Overall, automated extraction of DNA from milk matrix combined with ARMS-PCR is the most suitable technique to provide genetic characterization of the CSN2 gene on a large scale.

Grazing affects metabolic pattern of individual cow milk.

Effective traceability tools able to characterize milk from pasture are important to safeguard low-input farming systems, niche dairy products, and local traditions. The aims of the present study were to investigate the ability of proton nuclear magnetic resonance (1H NMR) spectroscopy to discriminate between milk produced from cows before and after the beginning of the grazing season, and to assess the effects of grazing on milk metabolites. The research trial involved a single alpine holding with 72 lactating cows. Individual milks were repeatedly sampled from the same animals before (i.e., d -3 and -1) and after (i.e., d 2, 3, 7, 10, and 14) the onset of the grazing period. One-dimensional 1H NMR spectra of milk extracts were collected through a Bruker spectrometer. Random forest discriminant analysis was applied to 1H NMR spectra to predict the period of collection for each sample. Data concerning the relative abundance of milk metabolites were analyzed through a linear mixed model, which included the fixed effects of period of sampling, cow breed, stage of lactation, and parity, and the random effect of cow nested within breed. The random forest model exhibited great accuracy (93.1%) in discriminating between samples collected on d -3, -1, 2, and 3 and those collected on d 7, 10, and 14. Univariate analysis performed on the 40 detected metabolites highlighted that milk samples from pasture had lower levels of 14 compounds (with fumarate being the most depressed metabolite) and greater levels of 15 compounds (with methanol and hippurate being the most elevated metabolites). Results indicate that milk 1H NMR spectra are promising to identify milk produced in different conditions. Also, our study highlights that grazing is associated with significant changes of milk metabolic profile, suggesting the potential use of several metabolites as indicators of farm management.

Genome-wide assessment of diversity and differentiation between original and modern Brown cattle populations.

Identifying genomic regions involved in the differences between breeds can provide information on genes that are under the influence of both artificial and natural selection. The aim of this study was to assess the genetic diversity and differentiation among four different Brown cattle populations (two original vs. two modern populations) and to characterize the distribution of runs of homozygosity (ROH) islands using the Illumina Bovine SNP50 BeadChip genotyping data. After quality control, 34 735 SNPs and 106 animals were retained for the analyses. Larger heterogeneity was highlighted for the original populations. Patterns of genetic differentiation, multidimensional scaling, and the neighboring joining tree distinguished the modern from the original populations. The FST -outlier identified several genes putatively involved in the genetic differentiation between the two groups, such as stature and growth, behavior, and adaptability to local environments. The ROH islands within both the original and the modern populations overlapped with QTL associated with relevant traits. In modern Brown (Brown Swiss and Italian Brown), ROH islands harbored candidate genes associated with milk production traits, in evident agreement with the artificial selection conducted to improve this trait in these populations. In original Brown (Original Braunvieh and Braunvieh), we identified candidate genes related with fat deposition, confirming that breeding strategies for the original Brown populations aimed to produce dual-purpose animals. Our study highlighted the presence of several genomic regions that vary between Brown populations, in line with their different breeding histories.

Short communication: Iodine content in bovine milk is lowly heritable and shows limited genetic variation.

Milk and dairy products are considered important sources of iodine in several countries. Despite this, there is a paucity of studies that have investigated sources of variation of milk iodine, especially on a large scale. So far, it is not clear if milk iodine content could be increased through breeding in dairy cattle. Recently, a mid-infrared spectroscopy prediction model has been developed for an indirect quantification of iodine content in cow milk, as it is a faster and less expensive method that allows the prediction at population level. The model has coefficient of determination and ratio of performance to deviation in external validation of 0.57 and 1.44, respectively, and it was used in the present study to predict the iodine content from historical milk spectral data to investigate phenotypic and genetic aspects in the Italian Holstein cattle. Based on the accuracy of the model, the prediction was interpreted as proxy for the real milk iodine concentration (IODP). The data set comprised 33,776 test-day records with IODP from 4,072 cows. Data of IODP were transformed through natural logarithm to achieve a normal distribution. The effect of parity, lactation stage, and month of sampling were investigated, and genetic parameters were estimated using a test-day repeatability animal model. Milk IODP decreased with parities and was the lowest in early lactation. Heritability of IODP was low (0.025) and it was positively genetically correlated with milk yield and negatively with fat content. Results suggested that it would be challenging to directly improve this trait through breeding strategies in dairy cattle, because IODP is mainly affected by temporary environmental factors and thus, cannot be easily improved through genetics. Although preliminary, findings of this study suggest that it would be more convenient to develop feeding and management strategies to drive milk iodine level than to put efforts and resources into breeding strategies. Further studies should validate IODP as an indicator trait of milk iodine content by improving reference data and estimating genetic correlation between predicted and measured values.

Short communication: Genetic aspects of milk urea nitrogen and new indicators of nitrogen efficiency in dairy cows.

Milk urea nitrogen (MUN), a trait routinely measured in the national milk recording system, is a useful indicator of nitrogen utilization efficiency of dairy cows, and selection for MUN and MUN-derived traits could be a valid strategy to produce better animals with regard to efficiency of nitrogen utilization. Therefore, the aim of the present study was to explore the genetic aspects of MUN and new potential indicators of nitrogen efficiency, namely ratios of protein to MUN, casein to MUN, and whey protein to MUN, in the Italian Brown Swiss population. A total of 153,175 test-day records of 10,827 cows in 500 herds were used for genetic analysis. Variance components and heritability of the investigated traits were estimated using single-trait repeatability animal models, whereas genetic and phenotypic correlations between the traits were estimated through bivariate repeatability animal models, including fixed effects of herd-test-date, stage of lactation, parity, calving year, and calving season, and the random effects of additive genetic animal, cow permanent environment, and the residual. Heritability estimates for MUN (0.20 ± 0.01) and the 3 new indicators of nitrogen utilization efficiency (0.15 ± 0.01 for protein-to-MUN and casein-to-MUN ratios, and 0.12 ± 0.01 for ratio of whey protein to MUN) suggested that additive genetic variation exists for these traits, and thus there is potential to select for greater organic nitrogen and lower inorganic nitrogen in milk. Genetic association between MUN and the 3 ratios was high (-0.87 ± 0.01) but not unity, suggesting that ratios could provide some further information beyond that provided by MUN with regard to efficiency of nitrogen utilization. Genetic trend of the investigated traits by year of birth of Brown Swiss sires showed how the selection applied in the last 30 yr has led to an increase of both quantity and quality of milk, and a decrease of somatic cell score and MUN. The inclusion of MUN in breeding programs could speed up the process of increasing organic nitrogen such as protein, which is useful for cheese-making, and reducing inorganic nitrogen (MUN) in milk.

A survey on sensor systems used in Italian dairy farms and comparison between performances of similar herds equipped or not equipped with sensors.

Sensor systems (SS) were developed over the last few decades to help dairy farmers manage their herds. Such systems can provide both data and alerts to several productive, behavioral, and physiological indicators on individual cows. Currently, there is still a lack of knowledge on both the proportion of dairy farms that invested in SS and type of SS installed. Additionally, it is still unclear whether the performances of herds equipped with SS differ from those of similar herds managed without any technological aid. Therefore, the aims of this study were (1) to provide an insight into SS spread among Italian dairy farms and (2) to analyze the performances of similar herds equipped or not equipped with SS. To reach the former goal, a large survey was carried out on 964 dairy farms in the northeast of Italy. Farmers were interviewed by the technicians of the regional breeders association to collect information on the type of SS installed on farms and the main parameters recorded. Overall, 42% of the surveyed farms had at least 1 SS, and most of them (72%) reared more than 50 cows. Sensors for measuring individual cow milk yield were the most prevalent type installed (39% of the surveyed farms), whereas only 15% of farms had SS for estrus detection. More sophisticated parameters, such as rumination, were automatically monitored in less than 5% of the farms. To reach the latter goal of the study, a subset of 100 Holstein dairy farms with similar characteristics was selected: half of them were equipped with SS for monitoring at least individual milk yield and estrus, and the other half were managed without any SS. Average herd productive and reproductive data from official test days over 3 yr were analyzed. The outcomes of the comparison showed that farms with SS had higher mature-equivalent milk production. Further clustering analysis of the same 100 farms partitioned them into 3 clusters based on herd productive and reproductive data. Results of the Chi-squared test showed that the proportion of farms equipped with SS was greater in the cluster with the best performance (e.g., higher milk yield and shorter calving interval). However, the presence of a few farms equipped with SS in the least productive cluster for the same parameters pointed out that although the installation of SS may support farmers in time- and labor-saving or in data recording, it is not a guarantee of better herd performance.

Technical note: Repeatability and reproducibility of curd yield and composition in a miniaturized coagulation model.

Miniaturized coagulation (MC) models have been proposed for the evaluation of curd yield (CY) in individual milk samples of different dairy species and breeds, and for the analysis of cheese microstructure and texture. It is still unclear if MC using less than 50 mL of milk is suitable to evaluate CY and chemical composition, and if preservative added to raw milk may interfere with MC process. Therefore, this study aimed at evaluating repeatability and reproducibility of CY, curd moisture, and fat and protein content on curd dry matter (DM) from MC trials using 40 g of milk. Miniaturized coagulations were performed by 3 different operators on 3 consecutive days, using raw milk (RM) and raw milk added with preservative (RMP). Repeatability of CY, calculated as relative standard deviation on 6 miniaturized curds obtained within a day by the same operator, was below 5% for MC carried out with both RM and RMP. The Horwitz ratio, which is the ratio between measured and expected reproducibility, highlighted good reproducibility for CY from RM and fair reproducibility for CY from RMP. The same ratio highlighted lower accuracies for curd moisture and fat and protein content on curd DM, especially for MC trials carried out with RMP. The z-test was performed to evaluate the similarity between curds manufactured with RM and RMP in terms of average yield and chemical composition; z-scores did not highlight significant differences between values obtained from MC carried out with RM and RMP. It can be concluded that preservative had negligible effects on MC, giving the opportunity to extend milk physical and chemical stability, to schedule laboratory trials on longer time span, and to broaden the sample size within a batch of analyses.

Estimation of maximum thermo-hygrometric index thresholds affecting milk production in Italian Brown Swiss cattle.

It is known that heat stress affects dairy cow performance in multiple ways: physiological, behavioral, reproductive, and productive. The aim of the present study was to determine if a threshold of temperature-humidity index (THI) exists for multiple milk production traits (milk yield, fat-corrected milk, protein and fat yield and percentage, energy-corrected milk, cheese production, and cheese yield) in Italian Brown Swiss dairy cows from the period 15 d before the day of the Italian Breeders Association test-day sampling. A 10-yr data set (2009-2018) containing 202,776 test-day records of 23,296 Brown Swiss cows was matched with the maximum THI. In all parities considered, no THI thresholds were observed for milk yield in Brown Swiss. In contrast, a THI threshold of 75 was identified for fat-corrected milk. No THI threshold was found for fat percentage, but fat yield showed the highest THI thresholds in cows of first and second parity. Protein yield and cheese production were affected by heat stress with average THI threshold of 74. The THI thresholds identified indicate that the Brown Swiss breed has higher thermal tolerance versus literature values reported for Holstein cows. As THI rises, Brown Swiss cows tend to produce the same volume of milk, but with a decreasing quality with regard to components. Further study is necessary to estimate the genetic component of heat tolerance, in Brown Swiss cattle, considering that the correct estimation of THI thresholds represents the first step to identify components that could be included in selection procedures.

Short communication: Genetic aspects of milk differential somatic cell count in Holstein cows: A preliminary analysis.

The aim of the present study was to assess genetic variation and heritability of a novel indicator of udder health, milk differential somatic cell count (DSCC), which represents the percentage of neutrophils plus lymphocytes in the total somatic cell count (SCC). Furthermore, we estimated genetic and phenotypic correlations of DSCC with other milk traits routinely measured in Italian Holstein cows. Besides DSCC, test-day data included milk yield, composition traits (i.e., fat, protein, casein, and lactose percentages), pH, milk urea nitrogen, and SCC. After editing, the final data set included 10,709 test-day records of 5,142 cows in 299 herds. Mean of DSCC was 62.07%, which means that macrophages were approximately 38% of total SCC. Comparing our results with the literature offered compelling evidence of the importance of acquiring information about the proportion of the different cell types in milk to better define the udder health status. In addition, our analysis revealed, for the first time, that DSCC is a heritable trait, and heritability (0.08 ± 0.02) was higher than that of traditional somatic cell score (0.04 ± 0.02). Nevertheless, heritability of DSCC is still low compared with milk yield and quality traits. Single-trait analysis within parity showed that DSCC was less heritable in primiparous than in multiparous cows, whereas bivariate analysis confirmed that DSCC and somatic cell score were 2 different traits, as their genetic and phenotypic correlations differed from unity. From a genetic point of view, the DSCC was positively weakly associated with milk yield, lactose percentage, and milk urea nitrogen, and negatively associated with pH. Our findings contributed to the understanding of the genetic background of DSCC and are a precursor to the potential use of DSCC in breeding programs to enhance cow resistance to mastitis. However, further research is needed to determine the weight this novel trait should receive in a selection program aimed to reduce udder health problems.

Genetic relationships of lactose and freezing point with minerals and coagulation traits predicted from milk mid-infrared spectra in Holstein cows.

The aim of the present study was to assess the relationships of lactose percentage (LP), lactose yield (LY), and freezing point (FRP) with minerals and coagulation properties predicted from mid-infrared spectra in bovine milk. To achieve this purpose, we analyzed 54,263 test-day records of 4,297 Holstein cows to compute (co)variance components with a linear repeatability animal model. Parity, stage of lactation, season of calving, and herd-test-date were included as fixed effects in the model, and additive genetic animal, within- and across-lactation permanent environment, and residual were included as random effects. Lactose percentage was more heritable (0.405 ± 0.027) than LY (0.121 ± 0.021) and FRP (0.132 ± 0.014). Heritabilities (± standard error) of predicted milk minerals varied from 0.375 ± 0.027 for Na to 0.531 ± 0.028 for P, and those of milk coagulation properties ranged from 0.348 ± 0.052 for rennet coagulation time to 0.430 ± 0.026 for curd firming time. Lactose percentage showed favorable (negative) genetic correlations with milk somatic cell score (SCS) and FRP, and it was almost uncorrelated with casein-related minerals (Ca and P) and coagulation properties. Moreover, LP was strongly correlated with Na (-0.783 ± 0.022), a mineral known to increase in the presence of intramammary infection (IMI) and high somatic cell count. Indeed, Na is the main osmotic replacer of lactose in mastitic milk when the blood-milk barrier is altered during IMI. Being strongly associated with milk yield, LY did not favorably correlate with coagulation properties, likely because of the negative correlation of this trait with protein and casein percentages. Milk FRP presented moderate and null genetic associations with Na and SCS, respectively. Results of the present study suggest that the moderate heritability of LP and its genetic correlations with IMI-related traits (Na and SCS) could be exploited for genetic selection against mastitis. Moreover, selection for LP would not impair milk coagulation characteristics or Ca and P content, which are important for cheesemaking.

Validation of a gold standard method for iodine quantification in raw and processed milk, and its variation in different dairy species.

Adequate milk consumption significantly contributes to meeting the human iodine recommended daily intake, which ranges from 70 µg/d for infants to 200 µg/d for lactating women. The fulfilment of iodine recommended daily intake is fundamental to prevent serious clinical diseases such as cretinism in infants and goiter in adults. In the present study iodine content was measured in raw and processed commercial cow milk, as well as in raw buffalo, goat, sheep, and donkey milk. Iodine extraction was based on 0.6% (vol/vol) ammonia, whereas iodine detection and quantification were carried out through an inductively coupled plasma mass spectrometer analyzer. Among processed commercial cow milk, partially skimmed pasteurized milk had the greatest iodine content (359.42 µg/kg) and raw milk the lowest (166.92 µg/kg). With regard to the other dairy species, the greatest iodine content was found in raw goat milk (575.42 µg/kg), followed by raw buffalo (229.82 µg/kg), sheep (192.64 µg/kg), and donkey milk (7.06 µg/kg). Repeatability of milk iodine content, calculated as relative standard deviation of 5 measurements within a day or operator, ranged from 0.96 to 1.84% and 0.72 to 1.16%, respectively. The overall reproducibility of milk iodine content, calculated as relative standard deviation of 45 measurements across 3 d of analyses and 3 operators, was 4.01%. These results underline the precision of the proposed analytical method for the determination of iodine content in milk.

Short communication: Association analysis of diacylglycerol acyltransferase (DGAT1) mutation on chromosome 14 for milk yield and composition traits, somatic cell score, and coagulation properties in Holstein bulls.

The aim of the present study was to determine the allele frequencies of the diacylglycerol acyltransferase (DGAT1) K232A mutation in Italian Holstein bulls and to estimate the effect of the mutation on milk yield, composition, somatic cell score, and coagulation traits (rennet coagulation time and curd firmness). For this purpose, 349 Italian Holstein bulls were genotyped for the DGAT1 mutation on chromosome 14. Association analysis was performed by regressing the number of copies for the K allele on the deregressed estimated breeding value of the individual. Breeding values were calculated using field data routinely collected in Northeast Italy. The frequencies of the AA, KA, and KK genotypes were 59.6, 32.1, and 8.3%, respectively, and the minor allele frequency (K variant) was 24.7%. The K allele was significantly associated with greater fat yield and fat, protein, and casein percentages and with reduced protein:fat ratio. The association between the DGAT1 mutation and somatic cell score was not significant, whereas a favorable association between presence of the K allele and milk coagulation properties was found. Results from the present study confirmed the effect of the diallelic DGAT1 polymorphism K232A on milk production traits and, for the first time, provided evidence that this mutation also affects milk coagulation properties in the Italian Holstein breed.

Alternative somatic cell count traits exploitable in genetic selection for mastitis resistance in Italian Holsteins.

The aim of the present study was to characterize alternative somatic cell count (SCC) traits that could be exploited in genetic selection for mastitis resistance. Data were from 66,407 first-parity Holsteins in 404 herds. Novel SCC traits included average somatic cell score (SCS, log-transformation of SCC) in early lactation (SCS_150), standard deviation of SCS of the entire lactation (SCS_SD), the presence of at least one test-day (TD) SCC >400,000 cells/mL in the lactation, and the ratio of number of TD SCC >400,000 cells/mL to total number of TD in the lactation. Novel traits and lactation-mean SCS (SCS_LM) were analyzed using linear mixed or logistic regression models, including month of calving, year of calving, number of TD, and milk yield as fixed effects, and herd and residual as random terms. A multitrait linear animal model was applied to a random subset of 152 herds (n = 22,695 cows) to assess heritability of and genetic correlations between SCC traits. Alternative SCC traits were affected by the environmental factors included in the model; in particular, results suggested a seasonal effect and a tendency toward an improvement of the udder health status in the last years. Association was also found between novel SCC traits and milk production. Alternative SCC traits exhibited coefficients of additive genetic variation that were similar to or larger than that of traditional SCS_LM. Heritability of novel SCC traits was smaller than heritability of SCS_LM (0.126 ± 0.014), ranging from 0.044 ± 0.008 (SCS_SD) to 0.087 ± 0.010 (SCS_150). Genetic correlations between SCC traits ranged from 0.217 ± 0.096 (SCS_150 and SCS_SD) to 0.969 ± 0.010 (SCS_LM and SCS_150). Alternative SCC traits exhibited additive genetic variation that is potentially exploitable in breeding programs of Italian Holstein population to improve resistance to mastitis.

Technical note: Development and validation of a new method for the quantification of soluble and micellar calcium, magnesium, and potassium in milk.

Milk mineral content is a key trait for its role in dairy processes such as cheese-making, its use as source of minerals for newborns, and for all traits involving salt-protein interactions. This study investigated a new method for measuring mineral partition between soluble and micellar fractions in bovine milk after rennet coagulation. A new whey dilution step was added to correct the quantification bias due to whey trapped in curd and excluded volume. Moreover, the proposed method allowed the quantification of the diffusible volume after milk coagulation. Milk mineral content and concentration in whey, and diluted whey were quantified by acid digestion and inductively coupled plasma optical emission spectrometry. The repeatability of the method for micellar Ca, Mg, and K was between 2.07 and 8.96%, whereas reproducibility ranged from 4.01 to 9.44%. Recovery of total milk minerals over 3 spiking levels ranged from 92 to 97%. The proposed method provided an accurate estimation of micellar and soluble minerals in milk, and curd diffusible volume.

Technical note: Development and validation of an HPLC method for the quantification of tocopherols in different types of commercial cow milk.

In the present study, a methanol-fluorescence-based HPLC method was validated for its use to quantify α-tocopherol and γ-tocopherol in raw milk, whole UHT milk, partially skimmed UHT milk, whole pasteurized milk, and partially skimmed pasteurized milk. Repeatability and reproducibility, calculated as relative standard deviation of 10 measurements within the same day and 30 measurements across 3 d, respectively, were always below 5% for both tocopherols concentrations and retention times. Recovery was assessed through 3 spiking levels and it ranged from 89 to 107%. The method was able to detect the expected declines in tocopherols in milk exposed to UHT or skimming treatments. Vitamin E, calculated as the sum of α-tocopherol and γ-tocopherol, was similar in whole pasteurized and raw milk, averaging 1.57 and 1.56 mg/L, respectively, followed by whole UHT (1.33 mg/L), partially skimmed pasteurized (0.77 mg/L), and partially skimmed UHT milk (0.61 mg/L).

Short communication: Phenotypic characterization of total antioxidant activity of buffalo, goat, and sheep milk.

Free radicals are reactive and unstable waste molecules produced by cells, responsible of damages and alteration on DNA, proteins, and fat. The daily intake of antioxidant compounds, acting against free radicals and their detrimental effects, is essential for human health. Milk contains several compounds with antioxidant activity, and the sum of their reducing potential blocking free radicals development is defined as total antioxidant activity (TAA). This novel trait has been described in literature both in individual and bulk cow milk, but there are no reports from other dairy species. Therefore, the present study aimed to investigate phenotypic variation of TAA in individual samples of buffalo (n = 105), goat (n = 112), and sheep (n = 198) milk. Total antioxidant activity was measured through a reference spectrophotometric method, and expressed as millimoles per liter of Trolox equivalents (TE). The greatest TAA was observed in sheep milk, averaging 7.78 mmol/L of TE and showing also the broadest phenotypic variation expressed as coefficient of variation (13.98%). Significantly lower TAA values were observed for buffalo (7.35 mmol/L of TE) and goat (6.80 mmol/L of TE) milk, with coefficients of variation of 8.18 and 8.47%, respectively. Total antioxidant activity exhibited weak correlations with milk yield and chemical composition. Phenotypic values of TAA presented in this study will be used to assess the ability of mid-infrared spectroscopy to predict this new trait and thus to collect data at the population level.